自噬和线粒体辅酶Q对急性脓毒症大鼠胰腺外分泌功能的影响  

Effect of autophagy and mitochondrial coenzyme Q on exocrine function of pancreas in rats with acute sepsis

作　　者：陈胜利[1] 黄锦达 曾其毅[1] 贾玉娥[1] 王金华[1] 

机构地区：[1]南方医科大学附属珠江医院儿科,广东广州510280

出　　处：《中华危重病急救医学》2015年第2期86-91,共6页Chinese Critical Care Medicine

基　　金：基金项目：国家自然科学基金（81272070）

摘　　要：目的 探讨自噬对急性脓毒症大鼠胰腺外分泌功能的影响，以及线粒体辅酶Q（Mito Q）是否通过自噬对急性脓毒症大鼠胰腺外分泌功能障碍起保护作用。方法 实验1：将30只雄性SD大鼠按随机数字表法分为3组，每组10只。所有大鼠腹腔注射脂多糖（LPS）10mg／kg，1h后分别经尾静脉注射自噬特异性抑制剂渥曼青霉素（Wortmannin）2mg／kg（LPS＋Wortmannin组）、MitoQ6．5μmol／kg（LPS＋MitoQ组）或等量生理盐水（LPS组）。观察LPS后12h内动物存活情况。实验2：将100只雄性SD大鼠按随机数字表法分为10组：对照4h、6h、12h组，LPS4h、6h、12h组，LPS＋Wortmannin4h组，Wortmannin4h组，LPS＋Mito06h组，MitoQ6h组，每组10只，制模及给药方法同实验1。在相应时间点留取大鼠血标本，用速率法测定血清淀粉酶含量；取胰腺组织，用酶联免疫吸附试验（ELISA）检测活性氧（ROS）含量，蛋白质免疫印迹试验（WesternBlot）检测自噬相关基因LC3蛋白表达，镜下观察胰腺组织病理学改变。结果①LPS＋Wortmannin组大鼠存活时间明显短于LPS组（h：7．50±0．64比11．90±0．13，X^2=19．847，P=0．001）；LPS＋MitoQ组大鼠存活时间与LPS组无差异（h：11．60±0．24比11．90±0．13，x。=1．055，P=0．137）。②LPS6h组、LPS＋Wortmannin4h组、LPS＋MitoQ6h组血淀粉酶含量明显高于同时间点对照组（U／L：2881．00±550．12比2099．20±249．57，3672．00±779．24比2081．36±245．18，2975．20±687．03比2099．20±249．57，均P〈0．05），LPS12h组血淀粉酶含量明显低于同时间点对照组（U／L：794．00±218．71比2086．80±261．75，P〈0．01）；LPS6h和12h组、LPS＋Wortmannin4h组、LPS＋MitoO6h组胰腺组织ROS含量均明显高于同时间点对照组（kU／L：3．18±1．06比1．78±0．37，3．63±1．08比1．85±0．41，3．14±0．98比1．65±0．34，3．17±1．03比1．78±0．37，均P〈0．05）。Objective To investigate the effects of autophagy on exocrine function of pancreas in rats with acute sepsis, and to determine whether the mitochondrial coenzyme Q （Mito Q） can prevent exocrine dysfunction of pancreas mediated by autophagy. Methods Experiment [ ： 30 Sprague-Dawley （SD） rats were randomly divided into three groups, with 10 rats in each group. All the rats were given lipopolysaecharide （LPS, 10 mg/kg） intraperitoneally, and Wortmannin （2 mg/kg）, the specific inhibitor of autophagy （LPS ＋ Wortmannin group）, Mito Q （6.5 ~mol/kg, LPS ＋ Mito Q group）, or the same volume of normal saline （LPS group） was respectively injected via the tail vein 1 hour later.Survival rate was assessed within 12 hours after LPS injection. Experiment II ： another 100 male SD rats were randomly divided into ten groups with 10 rats in each group： namely control 4, 6 and 12 hours groups, LPS 4, 6 and 12 hours groups, and LPS ＋ Wortmannin 4 hours group, Wortmannin 4 hours group, LPS ＋ Mito Q 6 hours group, and Mito Q 6 hours group. The protocols of model reproduction and drug administration were the same as in the experiment I . Blood samples were collected at each time point, and the amylase content was determined with the velocity method. The levels of reactive oxygen species （ROS） in the pancreases were measured with enzyme-linked immunosorbent assay （ELISA）. The expression of the autophagy-related protein LC3 was determined with Western Blot. The pathological changes in the pancreas were observed with microscopy. Results ①The survival time in the LPS ＋ Wortmannin group was significantly shorter than that in the LPS group （hours： 7.50 ± 0.64 vs. 11.90±0. 13, X^2= 19.847, P = 0.001 ）. There was no significant difference in the survival time between LPS ＋ Mito Q and LPS groups （hours： 11.60±0.24 vs. 11.90±0.13, X^2= 1.055, P = 0.137）. ② The serum amylase in the LPS 6 hours, LPS ＋ Wortmannin 4 hours, and LPS ＋ Mito Q 6 hours groups were significantly h

关 键 词：自噬 线粒体辅酶Q 胰腺外分泌 脓毒症 大鼠 

分 类 号：R459.7[医药卫生—急诊医学]